The work proposed here is designed to build on what we have accomplished thus far toward establishing an efficient procedure for producing transgenic zebra finches. Zebra finches are songbirds that learn their song by imitating that of an adult. The brain regions that mediate this behavior are highly modular and well defined anatomically;they are collectively referred to as the "song system". Within this system a key population of neurons encoding the song is replaced continuously in adulthood. The tight relationship between anatomy and behavior, together with the rare ability to continuously replace those cells that produce the behavior, have made the song system a very attractive model for studying the basic biology of learning and neuronal replacement. Several key resources have been and are presently being generated to facilitate the study of this system at ever more reductionistic levels: the genome of this species has been sequenced, cDNA libraries have been generated, sequenced and published, and a BAC library has been created and made available. These resources have the potential to help elucidate the molecular mechanisms and cellular properties that mediate song behavior. However, to take full advantage of these resources it has to be possible to manipulate gene expression in vivo, which we cannot yet do in zebra finches. Over the last year this laboratory has devoted its resources to the development of such a tool, and the methods we have employed have allowed us to overcome key obstacles and make significant progress. That progress is outlined here as well as the steps we will take to produce an efficient protocol for making transgenic songbirds. PUBLIC HEALTH RELEVANCE: Neurogenesis and neuronal replacement occur in the context of song learning in adult zebra finches. This project is about the production of transgenic zebra finches that will help us understand the mechanisms, function and therapeutic potential of neuronal replacement in adult brain.